The effect of G-CSF on lymphocyte subsets and CD34+ cells in allogeneic stem cell transplantation.

The effect of granulocyte colony-stimulating factor (G-CSF) on peripheral blood lymphocytes (PBL) and CD34+ cell frequency in the apheresis product has been determined in 25 healthy stem cell donors. Peripheral blood mononuclear cells (PBMNC) were collected after five days of G-CSF 10 microg/kg/day s.c., which was well tolerated. The median number of leukocytes increased eight-fold over that of pretreatment levels. Collection of PBMNC lasted a median of two (range, 1-3) days. The mean mononuclear cell (MNC) count and total lymphocyte percentage were 6.69 x 10(8)/kg and 59.08%, respectively, and the frequency of CD34+ cell expression was 2.1% in the apheresis product. The frequency of CD3+, CD4+, CD25+, NK and CD122+ cell expressions in mobilized PBMNC and PBL showed no significant difference. However, the frequency of CD8+, CD8+28+, CD3+DR+, CD19+, CD20+ and CD22+ B cells expression in the apheresis product increased significantly compared to steady-state PBL. In contrast, the frequency of the CD11 a+ and CD8+38+ cell expressions in the apheresis product was decreased compared to the steady-state PBL. The mean yield of CD34+ and CD3+ cells were 13.6 x 10(6) and 2.69 x 10(8)/kg of recipient body weight (RBW), respectively. Following allograft all patients engrafted with >0.5 x 10(9)/l neutrophil and < or = 20 x 10(9)/l platelets on a median of day 13 and 12, respectively. Nine patients had grade II-IV acute GVHD and chronic GVHD occurred in eight patients. Four patients died due to transplant-related complications. There was one late engraftment failure which occurred on the fifth month. Thirteen patients are still alive. In conclusion, these results indicate that administration of G-CSF at 10 microg/kg/day in normal donors alters the lymphocyte subsets and there are significant differences in the lymphocyte contents of the recipients before apheresis and in apheresis product.     

Peripheral blood stem cell apheresis for allogeneic transplants: Ibni Sina experience

BACKGROUND: The rate of utilizing peripheral blood stem cells (PBSC) as a source for allogeneic stem cells is growing rapidly. We aimed to demonstrate our 4 years experience as the largest apheresis center in Turkey and analyzed the content of the apheresis material. PATIENTS AND METHODS: From 1998 to the end of April 2002, 151 leukopheresis procedures were performed on 116 healthy donors (M/F:66/50) with a median age of 30 years (14-53). The HLA identical sibling donors received rhG-CSF 10 microg/kg/day sc. for 4 days and at the 5th day leukopheresis was started until collecting >4 x 10e6/kg CD34+ cells. Two times the donors' total blood volume was processed in 195 min (178-245) on continuous flow cell separators using peripheral venous access. RESULTS: Preapheresis WBC was 51.5 x 10e9/L (range, 13.11-91.3). Mono nuclear cell, CD34 and CD3 quantity of the harvest material were 5.35 x 10e8/kg (range, 0.45-23.46), 6.4 x 10e6/kg (range, 2.49-33.27) and 2.79 x 10e8/kg (range, 0.46-30.95), respectively. We were able to reach the target CD34 count after 1st cycle in 39% and 2nd cycle 61% of the procedures. In all donors with a peripheral blood CD34 count >80/mcl we succeeded to collect enough stem cells with only one leukopheresis. CONCLUSION: Collection of peripheral blood stem cells with continuous flow cell separators is well tolerated, with no mobilization failures or poor mobilizers. We collected high values of CD34+ cells (med. 6.4 x 10e6/kg) at the expense of high CD3+lymphocytes (med. 2.79 x 10e8/kg), which may increase the risk of acute and chronic GVHD after allogeneic hemapoietic cell transplantation.     

Large-volume leukapheresis using femoral venous access for harvesting peripheral blood stem cells with the Fenwal CS 3000 Plus from normal healthy donors: predictors of CD34+ cell yield and collection efficiency

The current paper reports on the predicting factors associated with satisfactory peripheral blood stem cell collection and the efficacy of large-volume leukapheresis (LVL) using femoral vein catheterization to harvest PBSCs with Fenwal CS 3000 Plus from normal healthy donors for allogeneic transplantation. A total of 113 apheresis procedures in 57 patients were performed. The median number of MNCs, CD3+ cells, and CD34+ cells harvested per apheresis was 5.3 x 10(8)/kg (range, 0.3-11.0 x 10(8)/kg), 3.0 x 10(8)/kg (range, 0.2-6.6 x 10(8)/kg), and 7.9 x 10(6)/kg (range, 0.1-188.9 x 10(6)/kg), respectively. The median collection efficiency of MNCs and CD34+ cells was 49.8% and 49.7%, respectively. A highly significant correlation was found between the collected CD34+ cell counts and the pre-apheresis WBC counts in the donors (P = 0.013), and between the collected CD34+ cell counts and the pre-apheresis peripheral blood (PB) CD34+ cell counts (P<0.001). Harvesting at least >4 x 10(6)/kg CD34+ cells from the 1st LVL was achieved in 44 (77.2%) out of 57 donors and in 19 (90.5%) out of 21 donors with a PB-CD34+ cell count of >40/microl. There was no significant difference in the harvested MNC and CD34+ cell counts between the 1st and 2nd apheresis. The catheter-related complications included catheter obstruction (n = 2) and hematoma at the insertion site (n = 3). Accordingly, LVL using femoral venous access for allogeneic PBSC collection from normal healthy donors would appear to be safe and effective.     

Donor age-related differences in PBPC mobilization with rHuG-CSF

BACKGROUND: Data on the administration of rHuG-CSF to normal donors <18 years old are very limited. STUDY DESIGN AND METHODS: The results of rHuG-CSF administration to 61 donors <18 years old (Group A) were retrospectively evaluated and compared with results from 353 donors > or = 18 years old (Group B) who are included in the Spanish National Donor Registry. The mean age (range) in Group A and B was 14 (1-17) and 38 (18-71) years, respectively (p<0.001). The mean dose of rHuG-CSF was 10 microg per kg per day (range, 9-16) during a mean of 5 days (range, 4-6). Central venous access was placed more frequently in younger donors (25% vs. 6%; p<0.001). RESULTS: The mean number of CD34+ cells collected was 7.6 and 6.9 x 10(6) per kg of donor's body weight in Group A and B, respectively. Fifty-six percent of Group A donors needed only one apheresis to achieve > or = 4 x 10(6) CD34+ cells per kg versus 39 percent of Group B donors (p = 0.01). Side effects were more common in Group B (71% vs. 41%; p<0.001). CONCLUSION: The administration of rHuG-CSF to donors <18 years old leads to CD34+ cell mobilization in a pattern similar to that observed in adults. Greater age was associated with a more frequent requirement for more than one apheresis to achieve a similar number of CD34+ cells.     

重组人G-CSF对健康供者外周造血干／祖细胞动员和采集效果的影响因素分析

应用重组人粒系集落刺激因子（rhG—CSF）对健康供者进行动员并采集造血干细胞用于异基因外周血造血干细胞移植已在临床广泛应用，本研究通过对影响外周干细胞动员和采集效果的多因素分析，进一步探讨最佳动员方案及采集时机。采取回顾性方法分析了431例健康供者外周血干细胞动员采集效果，并进一步分析了供者一般特征、rhG—CSF动员天数、每日皮下注射次数、剂量与采集效果的关系。结果表明：rhG—CSF在动员中平均应用剂量为5．7μg／（kg·d），平均采集1．7次，收获单个核细胞数平均为9．57×10^8／kg，CD34^＋细胞平均为4．91×10^6／kg。绝大多数供者不良反应轻微。多因素分析结果显示，采集效率主要与供者体重指数，采集天数相关。rhG—CSF动员第5天采集的供者，其MNC数、CD34^＋细胞数及第一次单采成功率均优于其他时间采集的供者。同时，本组供者应用rhG—CSF剂量较小且剂量范围较窄，rhG—CSF剂量不如采集时间对采集物质量的影响明显。结论：小剂量应用rhG—CSF动员并于第5天开始采集是健康供者造血干细胞动员的较理想方案。     

低剂量rhG-CSF对56例非血缘供者外周造血干细胞动员

本研究观察低剂量人重组粒细胞集落刺激因子(rhG-CSF)对非血缘健康供者的影响,探讨用于中华造血干细胞捐赠者资料库提供的非血缘健康供者外周造血干细胞动员方案。56例非血缘健康供者接受rhG-CSF 5μg/(kg.d)皮下注射,在动员第4、5两天或第5、6两天采集干细胞,观察动员效果及不良反应,检测动员前后血常规指标、CD3+、CD4+、CD8+和CD20+细胞比例;对采集物进行单个核细胞(MNC)和CD34+细胞计数;对所有供者随访至2006年5月31日。结果显示:在rhG-CSF动员过程中出现1级毒副作用(按WHO分级标准):腰背酸痛17.9%(10/56)、焦虑失眠8.9%(5/56)、疲乏4.5%(3/56)等,无需特殊处理,无需终止动员。第4、5两天采集和第5、6两天采集所得的MNC分别是(5.95±1.52)×108/kg和(7.19±2.12)×108/kg;CD34+细胞分别是(3.03±1.09)×106/kg和(7.92±2.50)×106/kg。血红蛋白水平、血小板量、CD3、CD4、CD8、CD20百分比动员前后无变化。结论:5μg/(kg.d)rhG-CSF用于非血缘健康供者的动员是安全而有效的。     

The impact of the CD34+ cell dose on engraftment in allogeneic peripheral blood stem cell transplantation

Forty-five patients who underwent allogeneic peripheral blood stem cell transplantation (PBSCT) were evaluated in order to investigate any relationship between CD34+ cell dose given and hematological recovery. Granulocyte counts > 1.0 x 10(9)/L and platelet > 50 x 10(9)/L were considered as hematological recovery. Three different regimens were used for mobilization, by adjusting the recombinant granulocyte colony stimulating factor (rhG-CSF, Roche) dose. The first group (n = 3), whose donors mobilized with 5 micrograms/kg/d s.c. rhG-CSF received a mean of 5.9 x 10(6)/kg (95% confidence interval for mean (CI); 2.4-9.3) CD34+ cells. The second group (n = 37), mobilized with 10 micrograms/kg/d s.c. rhG-CSF and the third group (n = 5) mobilized with 15 micrograms/kg/d s.c. rhG-CSF, received a mean of 5.7 x 10(6)/kg (95% CI; 4.6-6.75) and 6.56 x 10(6)/kg (95% CI; 4.57-8.55) CD34+ cells, respectively. CD34+ cell dose was 5.82 x 10(6)/kg (95% CI; 4.97-6.68) for all the patients. All patients received rhG-CSF from day +1 until attaining granulocyte count > 1.0 x 10(9)/L for three consecutive days. Median granulocyte and platelet engraftment days for the whole group was 15 (range; 11-44) and 14 (11-54) days respectively. There was a close correlation (r = -0.301, p < 0.05) between the CD34+ cell dose and granulocyte recovery for the whole group. When these analyses were performed separately within groups, this correlation was also found significant for the first group (r = -0.99, p < 0.05) for granulocyte recovery. On the contrary the same analysis did not reach significance for the other groups, nor for platelet recovery for the whole group (r = 0.039, p = 0.821). We calculated a minimum dose of 4 x 10(6)/kg CD34+ cells for a safe alloPBSCT. There was no difference between patients who received more than 5 x 10(6)/kg CD34+ cells, and those who received more than 2 x 10(6)/kg and less than 5 x 10(6)/kg CD34+ cells. In conclusion, we have demonstrated a correlation between the CD34+ cell dose given and faster hematological recovery for alloPBSCT patients.     

Collection of two consecutive neutrophil concentrates for transfusion from donors mobilized with glycosylated granulocyte-CSF plus dexamethasone

BACKGROUND: The objective of this study was to establish a mobilization and apheresis regimen for collection of two consecutive polymorphonuclear neutrophil (PMN) concentrates from the same donor. STUDY DESIGN AND METHODS: In this prospective study, 111 healthy unrelated volunteers underwent either one (Group 1, n = 57) or two consecutive granulocyte apheresis procedure (Group 2, n = 54) using the a cell separator (Spectra). Both Group 1 and 2 donors were initially mobilized with glycosylated G-CSF 6.0 micro g per kg (range, 5.2-7.0 micro g/kg) subcutaneously plus oral dexa-methasone (DXM, 8 mg) and underwent granulocyte apheresis (GA-1) 16 hours (range, 13-18 hr) after initial G-CSF+DXM. Group 2 donors were remobilized with a second DXM dose of 8 mg (n = 13), 4 mg (n = 15), 1.5 mg (n = 13), or none (n = 13), and a second apheresis (GA-2) was run 40 hours (range, 37-42 hr) after G-CSF+DXM administration and 12 hours after remobilization with DXM alone. RESULTS: Based on equivalent median preapheresis WBC and PMN counts of around 35 x 10(9) WBCs per L and 33 x 10(9) PMNs per L after initial mobilization the GA-1 yields were 85 x 10(9) PMNs per U (range, 34-150) in Group 1 and 75 x 10(9) PMNs per U (range, 35-135) in Group 2 (p = 0.14, NS). In Group 2, median preapheresis values of 19.6 x 10(9) WBCs per L (range, 9.5-37.0) and 16.6 x 10(9) PMNs per L (range, 8.8-34.8) were measured after remobilization and GA-2 yields of 49 x 10(9) WBCs per U (range, 26-113) and 42 x 10(9) PMNs per U (range, 21-95) were obtained. Borderline statistical differences in the GA-2 yields were observed from the remobilized donors: 8 mg: 36 x 10(9) PMNs per U (range, 23-60); 4 mg: 47 x 10(9) PMNs per U (range, 21-56) (p 相似文献   

Technical and safety aspects of blood and marrow transplantation using G-CSF mobilized family donors

An allogeneic transplantation programme using immunoselected blood progenitor and bone marrow CD34+ cells has been established. Thirteen healthy HLA-matched, MLC negative sibling donors received two doses of 5 micrograms kg-1 G-CSF (s.c. daily) for 5 days. On days 4 and 5, large-volume mononuclear cell aphereses were performed (COBE Spectra) and on day 5 one unit of autologous blood was obtained. Mononuclear cells were pooled and cryopreserved after CD34+ cell-immunoselection on day 5. Bone marrow (BM) of the same donors was procured under routine conditions 10-45 days later (median: 27 days). The final graft consisted of blood CD34+ cells with either complete BM (n = 5) or immunoselected BM CD34+ cells (n = 8). The present paper describes the progenitor cell mobilization and apheresis protocol and analyzes the cell loss by BM and peripheral blood progenitor cell (PBPC) donation. Considerably larger amounts of mononuclear cells (CD45+), T-lymphocytes (CD3+) and platelets were lost by the apheresis as compared to bone marrow without apparent immediate clinical consequences for the donors. Owing to cross-cellular contamination of the apheresis concentrate, blood platelet count (PC) significantly decreased (mean PC after the second apheresis 116 x 10 microL-1); furthermore on average 3.04 x 10(10) CD3+ cells were removed by two apheresis sessions. This loss did not lead to long-term total lymphocyte count changes (2370 microL-1 versus 1889 microL-1) as observed during the long-term follow-up of 7/13 donors (mean 290 days). Subjectively, the PBPC collections were better accepted than BM donations in all but one family donor.     

Mobilization factors of peripheral blood stem cells in healthy donors.

As a source of hematopoietic stem cells for transplantation, the use of peripheral blood stem cells (PBSCs) has become routine and comparable to that of the use of bone marrow. Recently, elderly patients with hematological malignancies also have been allowed to receive minitransplantations with nonmyeloablative conditioning regimens under sufficient PBSC infusion. As a result of these minitransplantations, elderly donors have been chosen increasingly from the siblings of elderly patients. We analyzed factors influencing the condition of CD34+ cells in the first days of collection in 49 healthy donors from July 1995 to January 2001. The median dose of recombinant human granulocyte colony-stimulating factor was 8 microg/kg/day (range 8 - 10) over 3 days. The target number of CD34+ cells used in this study was > or = 3 x 10(6) cells/kg of recipient body weight. The median apheresis volume was 12 L. Except for one 60 year old man, we obtained an adequate number of stem cells. In the regression analysis, a negative correlation was seen between donor age and the number of CD34+ cells/kg of recipient body weight per 12 L volume (Y = aX + b; a = -0.07507; b = 6.629996; r = -0.50985; p = 0.000252). Significantly higher apheresis results were obtained in donors younger than 45 years compared with donors 45 years old and older (p < 0.0227). There were no correlations among the number of CD34+ cells, donor body weight, and the number of leukocytes in peripheral blood on the first day of apheresis.     

MAG方案对恶性血液病患者造血干细胞动员作用的研究

目的　研究米托蒽醌 (MTZ)联合大剂量阿糖胞苷 (Ara C)、重组人粒细胞集落刺激因子(rhG CSF)组成MAG方案对恶性血液病患者外周血干细胞的动员作用。方法　 1995年 12月至2 0 0 3年 4月 ,采用MAG方案对 14例恶性淋巴瘤和 2 9例急性白血病患者外周血干细胞进行动员 ,其用量为MTZ 10mg/m2 ,第 2 ,3天 ;Ara C 2 g/m2 ,每 12h 1次 ,第 1,2天 ;rhG CSF 30 0 μg/d。首先用MA方案联合化疗 ,白细胞 <1.0× 10 9/L时开始用rhG CSF ,白细胞回升时用CS 30 0 0plus或CobeSpectra血细胞分离机采集外周血干细胞。结果　 14例恶性淋巴瘤患者除 1例外周血干细胞采集失败外 ,其余 13例均 1次性采集成功 ,所得单个核细胞 (MNC) (3.91± 2 .70 )× 10 8/kg ,CD34 细胞 (17.79± 12 .90 )× 10 6/kg。采集 2 9例急性白血病患者外周血干细胞平均 2 .13次 ,2 4例采集成功 ,5例采集失败 ,所得MNC (3.6 2± 2 .89)× 10 8/kg ,CD34 细胞 (7.37± 6 .6 0 )× 10 6/kg。rhG CSF平均使用时间为 7d。经MAG方案动员后 ,除 8例患者有胃肠道反应、14例患者骨髓抑制期合并感染外无明显不良反应 ,无动员相关死亡。MAG方案动员后进行微小残留病检测 ,部分病例转为阴性。结论　MAG方案在恶性淋巴瘤和急性白血病患者外周血干细胞动员中安全     

异基因造血干细胞移植健康供者外周血干细胞二次采集时机探讨

目的 探讨健康供者体内应用rhG-CSF初次采集造血于细胞以后二次外周血造血干细胞采集的时机.方法 38例二次采集外周血干细胞的健康供者皮下注射rhG-CSF 5 μg·kg-1·d-1,连用5 d,在第5、6天采集外周血移植物(A组);A组供者初次采集时的资料作为对照(B组);A组供者依据初次和二次采集的75%间隔时间分为C组(≤9个月,n=30)和D组(>9个月,n=8).应用流式细胞术检测各组供者外周血采集物中的淋巴细胞,CD3+、CD3+CD4+、CD3+CD8+、CD14+、CD34+细胞以及CD3+CD4-CD8-T细胞的数量.结果 A组供者外周血采集物中淋巴细胞CD3+CD8+(25.51×108)和CD34+细胞(0.51×108)的中位含量显著低于B组(31.55×108和0.70×108,P<0.05);C组供者CD3+CD8+(23.42×108)和CD34+细胞(0.42×108)的中位含量显著低于B组(P<0.05);D组供者淋巴细胞,CD3+、CD3+CD4+、CD3+CD8+、CD14+、CD34+细胞以及CD3+CD4-CD8-T细胞的中位含量与B组的差异无统计学意义(P>0.05).A、C和D三组采集物中CD4+与CD8+细胞的比值、单核细胞与CD3+细胞的比值以及CD3+CD4-CD8-T细胞与CD3+细胞的比值与B组的差异均无统计学意义(P>0.05).38名健康供者初次和二次采集的间隔时间与二次采集物中的CD34+细胞存在显著的相关性(r=0.357,P=0.028).结论 健康供者二次采集外周血造血干细胞的时机以初次采集的9个月后为宜,9个月内采集应适当增加循环血量以保证移植物中的免疫和造血组分能满足临床需要.     

PBPC mobilization with paclitaxel, ifosfamide, and G-CSF with or without amifostine: results of a prospective randomized trial

BACKGROUND: The impact of amifostine on PBPC mobilization with paclitaxel and ifosfamide plus G-CSF was assessed. STUDY DESIGN AND METHODS: Forty patients with a median age of 34 years (range, 19-53) who had germ cell tumor were evaluated for high-dose chemotherapy. Patients were randomly assigned to receive either a single 500-mg dose of amifostine (Group A, n = 20) or no amifostine (Group B, n = 20) before mobilization chemotherapy with paclitaxel (175 mg/m(2)) given over 3 hours and ifosfamide (5 g/m(2)) given over 24 hours (TI) on Day 1. G-CSF at 10 microg per kg per day was given subsequent to TI with or without amifostine from Day 3 until the end of leukapheresis procedures. RESULTS: In 2 (10%) of 20 patients receiving amifostine and 3 (15%) of 20 patients not receiving it, no PBPC separation was performed because of mobilization failure. No significant differences were observed in the study arms with regard to the time from chemotherapy until first PBPC collection or the number of apheresis procedures needed to harvest more than 2.5 x 10(6) CD34+ cells per kg. Furthermore, leukapheresis procedures yielded comparable doses of CD34+ cells per kg (3.4 x 10(6) vs. 3.6 x 10(6); p = 0.82), MNCs per kg (2.7 x 10(8) vs. 2.6 x 10(8); p = 0.18), and CFU-GM per kg (15.9 x 10(4) vs. 19.3 x 10(4); p = 0.20). Patients in Group A had higher numbers of circulating CD34+ cells on Day 10 (103.0/microL vs. 46.8/microL; p = 0.10) and on Day 11 (63.0/microL vs.14.3/microL; p = 0.04) than did patients in Group B. CONCLUSION: Administration of a single dose of amifostine before chemotherapy with TI mobilized higher numbers of CD34 cells in the circulation, but did not enhance the overall collection efficiency in the present trial.     

健康供者特征对rhG-CSF预激的骨髓采集物CD34+细胞和T细胞亚群的影响

目的 探讨健康供者特征对rhG-CSF预激的骨髓(G-BM)采集物造血和免疫细胞成分的影响.方法 150名健康供者体内应用rhG-CSF 5 μ·kg-1·d-1连续4～5 d,第4,5天采集骨髓和外周血,用流式细胞术测定G-BM采集物中的CD3+、CD3+CD4+、CD3+CD8+、CD14+、CD34+细胞以及CD3+CD4-CD8-调节性T细胞的数量,并分析供者性别、年龄、体重、是否妊娠等特征对G-BM成分的影响.结果 150名健康供者G-BM所含有核细胞,淋巴细胞,CD3+、CD3+CD4+、CD3+CD8+、CD14+、CD34+细胞以及CD3+CD4-CD8-调节性T细胞的中位值分别为31 050(12 200～58 100)、2122(506～6618)、1344(307～4791)、692(145～3038)、616(112～2575)、986(265～2958)、63(11～505)和83(9～390)/μl.年龄≤38岁的供者G-BM中的有核细胞、CD34+细胞以及CD3+ CD4-CD8-调节性T细胞的数量分别为33800(18600～57100)、76(22～505)和97(11～380)/μl,显著高于年龄＞38岁的供者[分别为28 000(12 200～58 100)、49(11～220)和65(9～389)/μl],P值分别为＜0.05,＜0.001和0.001.外周血单核细胞计数＞0.37×109 /L的供者G-BM中有核细胞的数量[33550(12 200～57 100)/μl]显著高于≤0.37×109 /L的供者[27 150(13 800～58 100)/μl](P=0.005).多因素分析显示年龄和外周血单核细胞是G-BM中有核细胞采集量的影响因素[HR值分别为0.41和2.87;P值分别为0.01和0.003];年龄是G-BM中CD34+细胞采集量的唯一影响因素(HR=0.26;P值=0.001);年龄还是G-BM中CD3+ CD4-CD8-调节性T细胞数量的影响凶素[HR值为0.31;P=0.001].结论 年龄是影响G-BM中有核细胞、CD34+细胞和CD3+ CD4-CD8-调节性T细胞采集数量的因素,年龄≤38岁的供者更易采集满足临床需要的有核细胞和CD34+细胞;外周血单核细胞＞0.37×109 /L的供者采集的G-BM含有较多数量的有核细胞.     

Allogeneic blood progenitor cell collection in normal donors after mobilization with filgrastim: the M.D. Anderson Cancer Center experience

BACKGROUND: Information on the safety and efficacy of allogeneic peripheral blood progenitor cell (PBPC) collection in filgrastim-mobilized normal donors is still limited. STUDY DESIGN AND METHODS: The PBPC donor database from a 42-month period (12/94-5/98) was reviewed for apheresis and clinical data related to PBPC donation. Normal PBPC donors received filgrastim (6 microg/kg subcutaneously every 12 hours) for 3 to 4 days and subsequently underwent daily leukapheresis. The target collection was > or =4 x 10(6)CD34+ cells per kg of recipient's body weight. RESULTS: A total of 350 donors were found to be evaluable. Their median age was 41 years (range, 4-79). Their median preapheresis white cell count was 42.8 x 10(9) per L (range, 18.3-91.6). Of these donors, 17 (5%) had inadequate peripheral venous access. Leukapheresis could not be completed because of apheresis-related adverse events in 2 donors (0.5%). Of the 324 donors evaluable for apheresis yield data, 221 (68%) reached the collection target with one leukapheresis. The median CD34+ cell dose collected (first leukapheresis) was 462 x 10(6) (range, 29-1463).The main adverse events related to filgrastim administration in donors evaluable for toxicity (n = 341) were bone pain (84%), headache (54%), fatigue (31%), and nausea (13%). These events were rated as moderate to severe (grade 2-3) by 171 (50%) of the donors. In 2 donors (0.5%), they prompted the discontinuation of filgrastim administration. CONCLUSION: PBPC apheresis for allogeneic transplantation is safe and well tolerated. It allows the collection of an "acceptable" PBPC dose in most normal donors with one leukapheresis, with minimal need for invasive procedures.     

Proteolytic enzyme levels are increased during granulocyte colony-stimulating factor-induced hematopoietic stem cell mobilization in human donors but do not predict the number of mobilized stem cells

Previous studies from our laboratory indicate that functional, mature neutrophils are essential for interleukin-8 (IL-8)-induced stem cell mobilization. To study a possible role of neutrophils in granulocyte colony-stimulating factor (G-CSF) induced hematopoietic mobilization, we assessed the number of circulating CD34+ cells in healthy allogeneic stem cell donors on days 3, 4, and 5 of mobilization for comparison with the number of peripheral blood neutrophils and the plasma levels of IL-8, Flt3 ligand (FL), matrix metalloproteinase-9 (MMP-9), and human neutrophil elastase (HNE). Thirty-seven of 45 donors required 1 day of apheresis to obtain 5 x 10(6) CD34+/kg recipient body weight (high responders), the remaining 8 donors required 1 extra day of apheresis on day 6 (low responders). On day 5, CD34+ numbers in the blood were significantly highe in high responders (116 x 10(3) +/- 10.4/ml) than in low responders (54.1 x 10(3) +/- 10.3, p < 0.001). In all donors, MMP-9 and HNE levels were increased compared to nonmobilized individuals, but in high responders, plasma MMP-9 levels on days 3-5 of mobilization were substantially higher than in low responders (p < or = 0.02 for MMP-9 and p = 0.89, p = 0.05 and p = 0.52 for HNE on days 3, 4, and 5, respectively). These results are in accordance with the hypothesis that neutrophils play a role in G-CSF-induced mobilization through the release of proteases such as MMP-9 and elastase. No change in plasma levels of IL-8 or Flt3 ligand was observed, suggesting that these cytokines do not play a role in stem cell mobilization. However, because stem cell numbers could not be predicted by proteolytic enzyme levels and/or neutrophil numbers, other undefined factors may be more important.     

The predictive value of white cell or CD34+ cell count in the peripheral blood for timing apheresis and maximizing yield

BACKGROUND: The collection of peripheral blood stem and progenitor cells (PBPCs) for transplantation can be time-consuming and expensive. Thus, the utility of counting CD34+ cells and white cells (WBCs) in the peripheral blood was evaluated as a predictor of CD34+ cell yield in the apheresis component. STUDY DESIGN AND METHODS: The WBC and CD34+ cell counts in the peripheral blood and the apheresis components from 216 collections were assessed. Sixty-three patients underwent mobilization with chemotherapy plus filgrastim, and 17 patients and 14 allogeneic PBPC donors did so with filgrastim alone. The relationship between the number of WBC and CD34+ cells in the peripheral blood and in the apheresis component was analyzed by using rank correlation and linear regression analysis. RESULTS: The correlation coefficient for CD34+ cells per liter of peripheral blood with CD34+ cell yield (x 10(6)/kg) was 0.87 (n = 216 collections). This correlation existed for many patient and collection variables. However, patients with acute myeloid leukemia had fewer CD34+ cells in the apheresis component at any level of peripheral blood CD34+ cell count. Components collected from patients with CD34+ cell counts below 10 x 10(6) per L in the peripheral blood contained a median of 0.75 x 10(6) CD34+ cells per kg. When the WBC count in the blood was below 5.0 x 10(9) per L, the median number of CD34+ cells in the peripheral blood was 5.6 x 10(6) per L (range, 1.0-15.5 x 10(6)/L). A very poor correlation was found between the WBC count in the blood and the CD34+ cell yield (p = 0.12, n = 158 collections). CONCLUSION: The number of CD34+ cells, but not WBCs, in the peripheral blood can be used as a predictor for timing of apheresis and estimating PBPC yield. This is a robust relationship not affected by a variety of patient and collection factors except the diagnosis of acute myeloid leukemia. Patients who undergo mobilization with chemotherapy and filgrastim also should undergo monitoring of peripheral blood CD34+ cell counts, beginning when the WBC count in the blood exceeds 1.0 to 5.0 x 10(9) per L.     

人重组粒细胞集落刺激因子动员后采集骨髓对健康供者外周血采集物成份的影响

本研究探讨健康供者体内应用人重组粒细胞集落刺激因子（G—CSF）后采集骨髓对外周血采集物成份的影响。62例健康供者皮下注射rhG—CSF5μg/（kg·d），连用5天，其中31例供者在第4、5天分别采集骨髓和外周血采集物（A组）；另31例供者于第4、5天均单采集外周血单个核细胞（B组）。应用流式细胞术检测两组供者外周血采集物中的淋巴细胞、CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD14^＋、CD34^＋细胞以及CD3^＋CD4^-CD8^-T细胞的数量。结果显示，A组供者每微升外周血采集物中单个核细胞中CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD14^＋、CD34^＋细胞以及CD3^＋CD4^-CD8^-T细胞的中位含量分别1．56×10^5μ1、8．56×10^4μl、6．12×10^4μl、3．38×10^4μl、2．27×10^4μl、3．83×10^4μl、744μl及3588μl，与B组的1．40×10^5μl、7．34×10^4μ1、5．32×10^4l、3．06×10^4μl、1．83×10^4μl、3．21×10^4μl、554μl及3120μl的差异无统计学意义（p〉0．05）；A组外周血采集物中CD4^＋细胞与CD8^＋细胞的比值[1．52（0．54—2．87）]、单核细胞与CD3^＋细胞的比值[0．57（0．15—1．64）]以及CD3^＋CD4^-CD8^-T细胞与CD3^＋细胞[0．064（0．018—0．673）]的比值与B组[1．68（0．31—3．35）]、[0．59（0．18—1．25）]、[0．063（0．021—0．136）]的差异均无统计学意义（P〉0．05）。结论：健康供者体内应用rhG—CSF后采集骨髓对外周血采集物成份无影响，对rhG—CSF动员的同一健康供者可单独采集骨髓、外周血采集物或同时采集两种采集物以满足临床移植的需要。     

Mobilization of peripheral blood stem cells with paclitaxel and rhG-CSF in high-risk breast cancer patients

Preclinical studies have demonstrated the rapid and efficient mobilization of hematopoietic peripheral blood stem cells (PBSC) in a mouse model using the combination of paclitaxel with recombinant human granulocyte colony-stimulating factor (rhG-CSF). On the basis of these results, a clinical trial was initiated using rhG-CSF with paclitaxel for PBSC mobilization in high-risk breast cancer patients. The mobilized PBSC were evaluated for CD34(+) cell number, mononuclear cell content, and clonogenic potential. One-hundred and seventeen breast cancer patients received paclitaxel (300 mg/m(2)) administered as a 24-h continuous intravenous infusion. Forty-eight hours after completing paclitaxel, rhG-CSF (5 microg/kg) was initiated and continued until completion of PBSC collection. Leukapheresis was initiated once the white blood cell count reached 1.0 x 10(9)/L. Each collection was evaluated for the numbers of mononuclear cells (MNC) and CD34(+) cells. Clonogenic potential was enumerated using colony-forming units-granulocyte-macrophage (CFU-GM) and burst-forming units-erythroid (BFU-E). Patients receiving paclitaxel with rhG-CSF mobilized a large number of mononuclear cells/apheresis (mean, 3.7 x 10(8); range, 3.3-4.1) and CD34(+) cells/apheresis (mean, 7.2 x 10(6); range, 6.1-8.4). The average number of leukophereses needed was 1.8 (mean, range 1.6-2.0). Colony growth was normal with 178.9 x 10(5) and 214.8 x 10(5) colonies counted in CFU-GM and BFU-E assays, respectively. Patients engrafted platelets and neutrophils on day 10 following transplantation. In conclusion, PBSC mobilization with paclitaxel and rhG-CSF results in a large number of mononuclear cells and CD34(+) cells with normal clonogenic potential. The cells engraft normally following high-dose chemotherapy and autologous stem cell transplantation in high-risk breast cancer patients. These results demonstrate that paclitaxel with rhG-CSF is an efficient mobilizing agent in high-risk breast cancer patients.